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. 2013 Feb 20;288(16):11047–11065. doi: 10.1074/jbc.M112.438473

FIGURE 6.

FIGURE 6.

Cooperative promoter activation by ELK1 and AR through the ELK1 binding element. A, HeLa cells were co-transfected with (ELK1)2-TATA-Luc and pSG5-AR or pSG5 vector control. Cells were treated with either vehicle or testosterone (10 nm), and luciferase activity was measured 48 h after transfection. B and C, HeLa cells were co-transfected with (ELK1)2-TATA-Luc and pCMV-ELK1 or pCMV vector control and pSG5-AR. Cells were treated with either vehicle (Veh) or testosterone (Test) (10 nm). 48 h after transfection, cells were harvested to measure ELK1 mRNA (B) or lysed to measure luciferase activity (C). D, HeLa cells were transfected and treated as described for C except that the promoter construct used was ARE-TATA-Luc. E, HeLa cells were transfected and treated as described for C except that the promoter construct used was ISRE-TATA-Luc. F, HeLa cells were co-transfected with (ELK1)2-TATA-Luc and ELK1 shRNA 1, ELK1 shRNA 2, or control shRNA plasmid and pSG5-AR. Cells were treated with testosterone (10 nm). 48 h after transfection, cells were harvested to measure ELK1 mRNA (left panel) or to measure luciferase activity (right panel). G, C4-2 cells were plated in hormone-depleted medium and nucleofected with either (ELK1)2-TATA-Luc (left panel) or PSA-promoter Luc (right panel) and treated with vehicle or R1881. 48 h after nucleofection, the cells were harvested for luciferase activity. H, C4-2 cells were nucleofected in hormone-depleted medium with (ELK1)2-TATA-Luc and AR shRNA, ELK1 shRNA, or both ELK1 and AR shRNAs. After 48 h of nucleofection, the cells were harvested for luciferase activity (left panel). The endogenous AR and ELK1 mRNA expression levels were measured by quantitative real time PCR (right panel). *, **, and §, p < 0.001. Error bars represent S.D. Ctrl, control; KD, knockdown; RLU, relative luciferase units.